Introduction
In this research, the identification of the role of resveratrol in the reduction of synthesis of prostaglandin and the free radicals can be evaluated. It can be stated that the reduction of microglial activation helps protect against numerous kinds of “brain injury”. Different types of factors can begin the “neuro-inflammatory responses”. This can be advantageous or it can also be very harmful to mammalian cells. The cells that are involved in neuro-inflammation can be stated as microglial cells. The release of the mediators can be possible by the “microglial cells”. Due to the contribution of the mediators, the death of neuronal cells in various conditions can be possible. This can be “cerebral ischemia” and neurodegenerative types of disorders. Resveratrol can be found to have potent antioxidant and anti-inflammatory properties in this study. With this, it can be demonstrated that “resveratrol” is a potent inhibitor of “COX-1 activity”. “Resveratrol” diminished neuronal cell passing as well as diminished the astrocytes as well as initiated “microglial cells”. The ability of this component to prevent neuronal death in cerebral ischemia appears to be based on its ability to scavenge free radicals. In the chain of events that result in tissue damage, neurodegeneration, and other forms of brain damage, microglial cells play a crucial role.
Figure 1: Impact of resveratrol on microglial cells
(Source: Self-created in MS word)
A. Main types of techniques with disadvantages, advantages, principles, and controls
The neuroprotective effects of resveratrol under “ischemic conditions” has been studied on a molecular level through the use of “hypoxic hippocampal”. With this, in vitro models with neuronal cultures and glucose has been studied. Resveratrol inhibited toxicity that is “nitric oxide-related” to culture “hippocampal neurons” in a manner that appears to be related to its antioxidant properties (Chimento et al. 2021). Similarly, the “phosphoinositide-3-kinase (PI3-K)/Akt pathway” was activated by resveratrol in “tissue-type hippocampal cultures” exposed to “hypoxia-glucose”. This is also effective for the reduction of cell death. The “neuroprotective effects” of resveratrol extend beyond “cerebral ischemia”. Within the rat models of traumatic brain injury, the natural compound is effective to reduce “oxidative stress” as well as damage volume. In a mouse model for “Huntington’s disease”, resveratrol prevented “oxidative stress”, “excitotoxicity”, and “behavioral changes” that can be caused by “kainic acid”. With this, it has recently been demonstrated that resveratrol stimulates the “proteasome-mediated intracellular degradation” of “amyloid peptides”.
The main techniques that are used in this research can be mentioned. The used techniques in this research are western blotting and the “RT-PCR” technique. Besides this, the identification of the different expressions of products like “COX enzymes”, and “LPS-treated mPGES-1” can be evaluated through the culture of the microglial cell (Terracina et al. 2022). The main principle in the research is the identification of the impact of resveratrol in the reduction of the formation of free radicals. The evaluation of the impact of resveratrol on cyclooxygenase activity can be identified with the help of the “RT-PCR” technique. This technique can be done based on the principles of “RT-PCR”. The impact of resveratrol is advantageous in the reduction of the rate of inflammation. But in the case of the surgical case within the people the reduction of the rate of taking resveratrol is important. The use of control in the research is very important for the proper identification of the impact of resveratrol in the reduction of the rate of inflammation within the microglial cells of the rat compared to untreated resveratrol cells.
B. Context of methods description
Primary cultures of mixed microglial cells of rats were made from the cerebral cortex of “one-day-old Sprague-Dawley” rats. After being harvested through centrifugation at 1000 rpm for ten minutes, the cell culture was suspended in “Dulbecco’s Modified Eagle’s Medium” which can be abbreviate as “DMEM”. Penicillin and 40 g/ml streptomycin are two more antibiotics. The inoculum was washed for removal of the cells that are “non-adherent” (Fuloria et al. 2022). Then “fresh medium” was added the following day. Immunofluorescence and cytology have yet to demonstrate that micro cultures are more than 98% pure. After collecting the supernatant and centrifuging it at 10,000 g for ten minutes, the prostaglandin E2 enzyme immunoassay (PGE2) was used to measure the concentration of PGE2 in the medium. An arachidonic etching measurement was carried out to ascertain whether resveratrol had any direct inhibitory effects on the activity of the COX-1 along with “COX-2” enzymes.
With the pre-incubation, the mouse primary microglial cells were plated in a culture plate containing 24 wells (Silva et al. 2019). The impact of resveratrol on “COX-1” microglial enzyme activity was examined. By LPS with a concentration of 10 ng/ml over 24 hours. The COX-1 isoform is communicated by unstimulated essential microglial cells. Due to this, the activity of the “COX-1” assay was carried out without the use of LPS before the experiment. Electrophoresis was used to separate the PCR products on an agarose gel containing 2%. The risk of “genomic DNA” contamination was reduced by avoiding the “reverse transcriptase” along with employing a primer for the gene during the amplification step of PCR. Then RNA samples were used. Under decreasing conditions, protein samples can be evaluated under “SDS-PAGE” due to “Western blotting” (Costa et al. 2021). The formation of 8-isoprostanes, which are regarded as an effective measure for the formation of free radicals, is the result of free radicals attacking arachidonic acid on membrane phospholipids. For thirty minutes, microglial cells were pretreated with varying concentrations of resveratrol.
C. Background of this particular figure
The provided figure can be evaluated and the background of provided photomicrographs can eb stated. In this photomicrograph, the expression of the different products like “beta-actin”, “COX-2″, “COX-1” as well as “and mPGES-1” is produced through the use of “The RT-PCR” technique. For the formation of this photomicrograph in this research, microglial cells were treated with LPS. The selected concentration of this lipopolysaccharide is 10 Nano grams per liter. This treatment of the LPS-contained microglial cells was done with the addition of resveratrol and without resveratrol addition (Alesci et al. 2022). This addition of resveratrol can be based on the numerous concentration. Based on the variation of the concentration of resveratrol the level of expression of the products can vary. The expression of the “mRNA” of the products is identified through this “RT-PCR” micrograph. “COX-1 mRNA” is expressed continuously in every circumstance that is low level and high-level concentration of “resveratrol”. In contrast, the untreated nature of “microglial cells” did not express “COX-2” or “mPGES-1”. But the presence of LPS significantly increased their expression. Resveratrol administration is effective for the reduction of the expression of “mPGES-1” but not the “COX-2” expression (Chedea et al. 2022). “RT-PCR” can be carried out after LPS incubation for four hours. 30 minutes before adding LPS to the culture medium, resveratrol was applied. The effect of resveratrol on the expression of mPGES-1 can be examined semi-quantitatively. In this research, the results are evaluated based on this figure in which the experiments of RT-PCR can be evaluated. The expression of beta-actin that is in the rat microglial cells can be presented as normalization. The size of the amplicon of mRNA for the product of the microglial cells can be identified through this RT-PCR technique. The figure based on which this background is evaluated can be provided below.
Figure 2: Micrograph of RT-PCR product of beta action, COX enzymes, and mPGES-1
(Source: Candelario-Jalil et al. 2007)
D. Explanation of the results systematically
Resveratrol has an inhibitory effect that is “dose-dependent” between 1 molar and 10 molar and has a strong inhibitory effect starting at 1 M on PGE2 synthesis. However, a higher dose of resveratrol was tested in this study (Lee et al. 2022). PGE2 remains at its original level because it does not further inhibit the formation of PGE2. As resveratrol is a very effective antioxidant, it could affect the production of “PGE2” by the activation of microglial cells. PGE2 biosynthesis is just marginally diminished at generally high dosages by the cancer prevention agents like alpha-tocopherol and its engineered analog Trolox C, dissimilar to resveratrol. PGE2 production did not decrease when these antioxidants were added to the micro-culture in lower doses.
After pre-incubating the cells for 30 minutes with resveratrol and LPS for 24 hours, PGE2 concentrations in the supernatant were measured in vitro. From 10 M to 50 M at the highest dose, resveratrol significantly inhibited “PGE2” production in microglial cells that is “LPS-stimulated” (Vladu et al. 2022). Taking care of conditions, RT-PCR investigation uncovered that the control microglial cells cannot communicate mRNA of “COX-2” along with “mPGES-1” mRNA. LPS significantly increased the expression of these enzymes within the culture medium. [Referred to Appendix 1]
Potential impacts of “SC-560” on “mPGES-1” coupling with LPS obstruction, which might assist with understanding whether the impact of resveratrol on mPGES-1 decrease is interceded by the COX-1 part or not. SC-560 and VAS COX-1 inhibition did not alter the expression of “mPGES-1” in primary microglial cells that are “LPS-activated”. Some issues require additional discussion. The ability of resveratrol to prevent the action of “COX-1 peroxidase” is one of its fine pharmacological effects.
E. Discussion and interpretation depend on the key findings
Resveratrol is one of the most promising molecules because new research demonstrates its potent ability to prevent post-ischemic or hypoxic brain damage and trauma damage (Gal et al. 2021). PGE2 synthesis and the production of free radicals that cause oxidative stress are two of the most significant processes that contribute to the negative effects of increased COX activity during neuro-inflammation. In the pathological processes that are associated with brain injury, neuritis plays a significant role. In particular, brain injury is primarily caused by the activation of microglia and the subsequent production of “inflammatory cytokines”, “reactive oxygen species (ROS)”, and “prostaglandins” (Salehi et al. 2018). Post-ischemic brain injury can be avoided by either increasing or decreasing COX-2 activity. The production of 8-iso-PGF2, a reliable biomarker for the formation of free radicals, was significantly inhibited at low levels of resveratrol. The main terminal combination for the PGE2 blend in actuated microglia, mPGES-1, was altogether diminished by resveratrol because of LPS. The impact of Resveratrol within the reduced expression of mPGES-1are mediated by the COX-1 mechanism. [Referred to Appendix 2]
“COX-1 isoform” is a significant free radical in microglia and is involved in the synthesis of PGE2 that is induced by LPS with adding resveratrol (Dull et al. 2019). This finding provided evidence that “COX-1 microglial activity” is a very effective free radical within “neuro-inflammation”. SC-560 and VAS COX-1 inhibition did not alter the expression of “mPGES-1” in primary microglial cells that are LPS-activated. Microglia continuously expressed COX-1 at rest, and LPS did not raise its expression. Reduced COX activity and the production of PGE2 are both aided by the capacity of resveratrol to maintain a minuscule redox state in these cells. This finding is that additional antioxidants are Trolox C, as well as alpha-tocopherol, which can similarly inhibit the production of PGE2 backs this up. Antioxidants have been shown to regulate micro-activation (Candelario-Jalil et al. 2007). The ability of resveratrol to suppress mPGES-1 expression in microglia is activated. Importantly, resveratrol reduced the expression of “mPGES-1” but not “COX-2”.
F. Future steps and key conclusion
From the overall demonstration of the essay, it can be stated that the neuroprotective effects of resveratrol may be supported by a significant decrease in PGE2 and the production of free radicals by activated microglia. This research lends credence to the idea that resveratrol could be used as a treatment to lessen the activation of microglia in brain injuries of all kinds. As resveratrol is very beneficial for the reduction of inflammation it can be used for the extension of life span within people. The effectiveness of this resveratrol is identified within the rat cells to use its effectiveness within the mammalian cells. Due ot the most effective impact of resveratrol this can be used as medication. The aging process of mammalian cells can be slowed down by us of this component. With this, the blood flow of the cerebral cortex can be improved by the use of resveratrol.
Reference list
Journals
Alesci, A., Nicosia, N., Fumia, A., Giorgianni, F., Santini, A. and Cicero, N., 2022. Resveratrol and immune cells: A link to improve human health. Molecules, 27(2), p.424.
Candelario-Jalil, E., de Oliveira, A.C.P., Gräf, S., Bhatia, H.S., Hüll, M., Muñoz, E. and Fiebich, B.L., 2007. Resveratrol potently reduces prostaglandin E2production and free radical formation in lipopolysaccharide-activated primary rat microglia. Journal of neuroinflammation, 4(1), pp.1-12.
Chedea, V.S., Macovei, Ș.O., Bocsan, I.C., Măgureanu, D.C., Levai, A.M., Buzoianu, A.D. and Pop, R.M., 2022. Grape Pomace Polyphenols as a Source of Compounds for Management of Oxidative Stress and Inflammation—A Possible Alternative for Non-Steroidal Anti-Inflammatory Drugs?. Molecules, 27(20), p.6826.
Chimento, A., Santarsiero, A., Iacopetta, D., Ceramella, J., De Luca, A., Infantino, V., Parisi, O.I., Avena, P., Bonomo, M.G., Saturnino, C. and Sinicropi, M.S., 2021. A phenylacetamide resveratrol derivative exerts inhibitory effects on breast cancer cell growth. International journal of molecular sciences, 22(10), p.5255.
Costa, T.J., Barros, P.R., Arce, C., Santos, J.D., da Silva-Neto, J., Egea, G., Dantas, A.P., Tostes, R.C. and Jimenez-Altayo, F., 2021. The homeostatic role of hydrogen peroxide, superoxide anion and nitric oxide in the vasculature. Free Radical Biology and Medicine, 162, pp.615-635.
Dull, A.M., Moga, M.A., Dimienescu, O.G., Sechel, G., Burtea, V. and Anastasiu, C.V., 2019. Therapeutic approaches of resveratrol on endometriosis via anti-inflammatory and anti-angiogenic pathways. Molecules, 24(4), p.667.
Fuloria, S., Sekar, M., Khattulanuar, F.S., Gan, S.H., Rani, N.N.I.M., Ravi, S., Subramaniyan, V., Jeyabalan, S., Begum, M.Y., Chidambaram, K. and Sathasivam, K.V., 2022. Chemistry, Biosynthesis and Pharmacology of Viniferin: Potential Resveratrol-Derived Molecules for New Drug Discovery, Development and Therapy. Molecules, 27(16), p.5072.
Gal, R., Deres, L., Toth, K., Halmosi, R. and Habon, T., 2021. The effect of resveratrol on the cardiovascular system from molecular mechanisms to clinical results. International Journal of Molecular Sciences, 22(18), p.10152.
Lee, I.T., Lin, H.C., Huang, T.H., Tseng, C.N., Cheng, H.T., Huang, W.C. and Cheng, C.Y., 2022. Anti-Inflammatory Effect of Resveratrol Derivatives via the Downregulation of Oxidative-Stress-Dependent and c-Src Transactivation EGFR Pathways on Rat Mesangial Cells. Antioxidants, 11(5), p.835.
Salehi, B., Mishra, A.P., Nigam, M., Sener, B., Kilic, M., Sharifi-Rad, M., Fokou, P.V.T., Martins, N. and Sharifi-Rad, J., 2018. Resveratrol: A double-edged sword in health benefits. Biomedicines, 6(3), p.91.
Silva, P., Sureda, A., Tur, J.A., Andreoletti, P., Cherkaoui-Malki, M. and Latruffe, N., 2019. How efficient is resveratrol as an antioxidant of the Mediterranean diet, towards alterations during the aging process?. Free radical research, 53(sup1), pp.1101-1112.
Terracina, S., Petrella, C., Francati, S., Lucarelli, M., Barbato, C., Minni, A., Ralli, M., Greco, A., Tarani, L., Fiore, M. and Ferraguti, G., 2022. Antioxidant Intervention to Improve Cognition in the Aging Brain: The Example of Hydroxytyrosol and Resveratrol. International Journal of Molecular Sciences, 23(24), p.15674.
Vladu, A.F., Ficai, D., Ene, A.G. and Ficai, A., 2022. Combination Therapy Using Polyphenols: An Efficient Way to Improve Antitumoral Activity and Reduce Resista
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